Essential oil is one type of oil produced from various parts of plants, form by thick liquid at room temperature but easily evaporate, until confer the distinctive aroma. The essential oil of citrus peel used in pharmacological world, one of them as an antibacterial compound. This research was experimental using Randomized Complete Random Design (RAL) which was then analyzed using SPSS method of one way anova. The results show that: Fhit ≥ Ftab then the treatment of variations concentration of essential oils have an effect, so (accept H1, reject H0). The essential oil of citrus peel is obtained from separated by water-steam distillation method. Water-steam distillation method has the advantage that the oil produced more pure, simple and environmentally friendly because does not involve dangerous solvents. The resulting yield is 0.1%. The result of identification of Gas Chromatography-Mass Spectroscopy (KG-SM) shows that the essential oil of citrus fruits (Citrus sinensis L.) has a component of dl-limonene compound is 99.78% and Octamethyl-Cyclotetrasiloxane which is a hydrocarbon compound is 0.22% . The essential oil of sweet orange peel (Citrus sinensis L.) obtained with variation concentrations are (0.25, 2, 10, 100, and 500 mg / mL). The antibacterial activity test was performed using disc diffusion method giving effect to test bacteria form inhibitor zone around the paper disc with the average diameter of inhibited zone respectively 2; 6.7; 7.5; 8.1; and 10.9 mm for Staphylococcus aureus and 0; 0; 0; 2.4; and 2.9 for Escherichia coli.